Untreated mung bean hypocotyls produced very little C(2)H(4) but, upon treatment with 10 millimolar Cu(2+) or 10 millimolar Cu(2+) + 10 millimolar Ca(2+), C(2)H(4) production increased 20- and 40-fold, respectively, within 6 hours. This increase in C(2)H(4) production was preceded and paralleled by an increase in 1-aminocyclopropanecarboxylic acid (ACC) content, but the level of S-adenosylmethionine (SAM) was unaffected, suggesting that the conversion of SAM to ACC is a key reaction in the production of wound-induced C(2)H(4). This view was further supported by the observation that application of aminoethoxyvinylglycine, a known inhibitor of the conversion of SAM to ACC, eliminated the increases in ACC formation and in C(2)H(4) production. A significant increase in C(2)H(4) production was observed in the albedo tissue of orange in response to excision, and it was paralleled by an increase in ACC content. In columella tissue of unripe green tomato fruit, massive increases in the C(2)H(4) production rate (from 0 to 12 nanoliters per gram per hour), in ACC content (from 0.05 to 12 nmoles per gram), and in ACC synthase activity (from 0 to 6.4 units per milligram protein) occurred during the 9-hour incubation period following excision. Infiltration with 0.1 millimolar cycloheximide, an inhibitor of protein synthesis, completely blocked wound-induced C(2)H(4) production, ACC formation, and development of ACC synthase activity. These data indicate that wounding induces the synthesis of ACC synthase, which is the rate-controlling enzyme in the pathway of C(2)H(4) biosynthesis and, thereby, causes accumulation of ACC and increase in C(2)H(4) production.